Cordyceps militaris activity RT-PCR detection primers and detection method

Abstract

The invention discloses cordyceps militaris activity RT-PCR detection primers and a detection method, and belongs to the technical field of molecular detection and identification. The primers have nucleotide sequences as shown in SEQ ID NO.1 and SEQ ID NO.2. By adopting the specific primers SEQ ID NO.1 and SEQ ID NO.2, a catalase gene fragment of cordyceps militaris can be effectively amplified, and the fragment is 341 bp in size and can be used for RT-PCR detection. 2-delta delta CT statistical analysis is adopted for detection results, compared with a control strain, sample value / control value is used as a judgment standard, when the ratio is larger than or equal to 1, the detected strain can be used for production; when the ratio is less than 1 and greater than 0.1, the detected strainhas weak activity and should be used with caution; and when the ratio is less than 1 / 10, the detected strain basically cannot be used for production of cordyceps militaris and cordyceps flowers. The detection result can be used to predict relative activity of cordyceps militaris, and provide technical basis for screening strains in production.

Description

technical field [0001] The invention relates to a RT-PCR detection primer and a detection method for the activity of Cordyceps militaris, belonging to the technical field of molecular detection and identification. Background technique [0002] Cordyceps militaris (L.) Link is a fungus belonging to the Ascomycotina subphylum, which is isolated from wild Cordyceps militaris. It can be used to produce Cordyceps militaris fruiting bodies and silkworm Cordyceps militaris. The product has been proven to have multiple functions. It is currently produced in some Asian countries such as China and South Korea. With the maturity of production technology and the expansion of market scale, there are many problems restricting the Cordyceps industry. Among them, the instability of strain activity affects production is an important reason. Many production enterprises have lost production due to the degradation of strains and increased production risks. Shrestha, B. et al. believed that whe...

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Problems solved by technology

The earlier research on the difference in the activity of Cordyceps strains was to use RAPD to compare the strains with better activity and the strains with poorer activity. However, this method has many shortcomings and is not suitable for application, but it clarifies the mutation and expression function of some genes. to weaken

Xiong, C., etc. used transgenic technology to enhance the antioxidant capacity of Cordyceps militaris to delay the denaturation of Cordyceps militaris; Lin Qingquan, etc. judged the activity of the strain according to the acidity and alkalinity of the strain product, and believed that the strain with strong activity secreted acidic substances, and the strain with weak activity The secretion ability of the strain is weakened, the acidity is reduced, and the activity of the strain can be judged by the indicator. He also studied the method of dehydrogenase activity to judge the activity of the strain; Shanghai Academy of Agricultural Sciences reported PCR to judge whether the strain is Cordyceps fungus. Activity determination

At present, there is no rapid and effective method for detecting the activity of Cordyceps militaris

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